Growth and Production of Volatile Compounds of Yarrow (Achillea Millefolium L.) Under Different Irrigation Depths

Anais da Academia Brasileira de Ciências (2018) 90(4): 3901-3910 (Annals of the Brazilian Academy of Sciences) Printed version ISSN 0001-3765 / Online version ISSN 1678-2690 http://dx.doi.org/10.1590/0001-3765201820180092 www.scielo.br/aabc | www.fb.com/aabcjournal Growth and production of volatile compounds of yarrow (Achillea millefolium L.) under different irrigation depths IVAN C.A. ALVARENGA1, FERNANDA V. PACHECO1, AMAURI A. ALVARENGA2, SUZAN K.V. BERTOLUCCI1 and JOSÉ EDUARDO B.P. PINTO1 1Laboratório de Cultura de Tecidos e Plantas Medicinais, Departamento de Agricultura, Universidade Federal de Lavras, Caixa Postal 3037, 37200-000 Lavras, MG, Brazil 2Laboratório de Crescimento e Desenvolvimento de Plantas, Departmento de Fisiologia Vegetal, Setor de Biologia, Universidade Federal de Lavras, Caixa Postal 3037, 37200-000 Lavras, MG, Brazil Manuscript received on January 29, 2018; accepted for publication on July 11, 2018 ABSTRACT Yarrow (Achillea millefolium L., Asteraceae) is an important medicinal plant used worldwide for its medicinal properties such as the analgesic, antioxidant and anti-inflammatory ones. The aim of this study was to evaluate the growth and production of photosynthetic pigments and of volatile constituents of Achillea millefolium L. under different irrigation depths. The treatments were the application of 55, 110, 220, 440 and 880 mm of water for a period of 110 days. Data were submitted to polynomial regression analysis at 5% probability, while the volatile constituents were analyzed by standard deviation. Different irrigation depths provided quadratic growth responses being the highest dry matter production at the depth of 440 mm. The contents of chlorophyll a, b, total and carotenoids were higher at the lower depth tested (55 mm). The major volatile compounds identified were sabinene, 1,8-cineol, borneol and β-caryophyllene. Increased water availability reduced the complexity of the volatile fraction of essential oil. Thus, it is recommended that the species be cultivated at 440 mm irrigation depth to have a higher production of dry matter and lower variation in the volatile profile of the essential oil. Key words: water stress, 1,8 cineol, chlorophyll, terpenes. INTRODUCTION in the cosmetics industry, skin care products and aromatherapy (Mohammadhosseini et al. Yarrow (Achillea millefolium L., Asteraceae) is a 2017). This essential oil presents great diversity perennial herb, native to Europe and growing in of chemical compounds, and caryophyllene, different countries. The species produces essential sabinene, γ-terpinene, borneol and chamazulene are oil in its leaves and inflorescences, and is used as mentioned as majoritarian (Kindlovits and Németh an analgesic, antioxidant, anti-inflammatory and is 2012). However, the chemical composition of its gastroprotective (Judzentiene and Mokute 2010). essential oil varies due to different factors. For In addition, its essential oil is used commercially example, Pinto et al. (2014a) observed significant Correspondence to: Ivan Caldeira Almeida Alvarenga changes in the essential oil compounds of yarrow E-mail: [email protected] when cultivated under different light conditions; An Acad Bras Cienc (2018) 90 (4) 3902 IVAN C.A. ALVARENGA et al. the same response was observed when there was condition of less water availability (Khorasaninejad lack of macro and microelements in a hydroponic et al. 2011). Thus, it can be observed that water system (Alvarenga et al. 2015). availability influences both the essential oil content Irrigated cultivation allows medicinal plants and its quality, yielding significant changes in to be grown year-round, thus providing the the terpenic compounds. In addition, the proper continuous supply of raw material. However, irrigation management combines the production of the plant productivity response as a function of vegetable matter with the contents of secondary water availability is specific, since it is related to metabolites (Alvarenga et al. 2012). the genetic characteristics of the plant, nutrient Due to the influence of water availability on availability, and the ability of the plant to allocate plant production, and lack of knowledge of the photoassimilates (Marschner et al. 1996). In response of yarrow to irrigation, the objective of most cases, pigments such as chlorophylls and this study was to evaluate the growth, production of carotenoids are rapidly degraded under water photosynthetic pigments and of volatile constituents limiting conditions; however, they can also be of Achillea millefolium L. under different irrigation accumulated depending on the species (Anjum et depths. al. 2011, Manivannan et al. 2007). The species Lippia sidoides Cham., cultivated MATERIALS AND METHODS under water limitation, for example, showed a reduction in the production of dry matter and The experiment was conducted in a greenhouse, in essential oil (Alvarenga et al. 2012). However, the sector of Plant Physiology at the Department the moderate water limitation in a cultivation of of Biology of the Universidade Federal de Lavras lemongrass (Cymbopogon citratus (DC.) Stapf) (UFLA). A species specialist identified the plants increased the production of essential oil (Pinto et and deposited the exsiccatae in the Bioscience al. 2014b). For basil (Ocimum basilicum L.) and tea Institute of the Universidade Federal do Rio Grande tree (Melaleuca alternifolia Chen.), intermediate do Sul, Brazil, with register number ICN 187014. water depths have proved ideal for the production Drainage microlysimeters with capacity of of dry matter (Silva et al. 2012, Pravuschi et al. 15L and mean diameter of 25 cm were used. They 2010). were filled with substrate composed of soil and Therefore, water availability for the production sand in the ratio of 2:1, and fertilized with NPK 3 of medicinal plants alters their growth and the 4-14-8 in the ratio of 2 kg per m of substrate. The synthesis of secondary compounds. However, it physicochemical characteristics of the soil were is necessary to understand how the availability analyzed at the Laboratory of Analysis of Soil of -3 of water changes the plant metabolism, because UFLA, and were: pH: 5.4; P: 4.13 mg dm ; K: -3 -3 alterations in the levels of active constituents can 73.32 mg dm , Ca: 2.30 cmolc dm , Mg: 0.30 -3 -3 compromise the parameters of quality, efficacy and cmolc dm , Al: 0.10 cmolc dm , H + Al: 2.90 -3 safety of the plant material (Baghalian et al. 2008). cmolc dm ; V: 49.00%; organic matter: 2.10 dag Misra and Srivastava (2000) observed significant kg-1, Clay: 70.00 dag kg-1; Silt: 16.00 dag kg-1 and increases in menthol and menthone levels for Sand: 14.00 dag kg-1. Mentha arvensis L. under water limitation (field To start the experiment, the microlysimeters moisture capacity of 10%). On the other hand, for were saturated with water, and then covered with Mentha piperita L., menthol content decreased plastic to prevent evaporation to allow the substrate by 36% and menthone content by 50%, under the to have 100% of field capacity. By the time water An Acad Bras Cienc (2018) 90 (4) THE INFLUENCE OF WATER ON THE PRODUCTION OF VOLATILES 3903 percolation ceased, the planting of seedlings was homogenized with 30 mL of 80% acetone (v/v). carried out. The contents of chlorophyll a, chlorophyll b and The seedlings (derived from cuttings) were carotenoids were calculated with the equations produced in expanded polypropylene trays filled proposed by Lichtenthaler and Buschmann (2001) with commercial substrate, and kept in intermittent from the absorbance readings at 663.2 nm, 646.8 mist chamber. After sixty days, with 10 cm of nm and 470 nm, respectively. All procedures height and four expanded leaves, the seedlings were performed in the dark and the readings were were transplanted to the microlysimeters. performed in triplicates. The treatments consisted of application of The constituents of the volatiles were analyzed five irrigation depths: 55, 110, 220, 440 and 880 from 40 mg of dry matter of leaves, and four mm, distributed in daily applications during the composite samples per treatment were utilized. growing period. For each treatment, five replicates Dried leaves were placed in a 20 mL headspace were used with a plant per repetition, in a total of vial, sealed with septum silicone / PTFE. The 25 plants, distributed in a completely randomized analyses were performed with the “headspace” design. Plants from which samples were taken technique in a Gas Chromatograph coupled to a for chlorophyll analysis were not accounted for Mass Spectrophotometer (GC/MS: Agilent® 7890 assessment of the production of dry matter of the A / Agilent® MSD 5975C, HP-5MS column). After aerial part and total dry matter, being used for optimization, the following operating conditions analysis of the other variables. were established: Sample incubation temperature of The average temperature during the experiment 100 °C for 30 min, syringe temperature of 110 °C. The was of 22.08 °C, with average maximum volume of 500 µL of the vapor phase was injected temperature of 30.91 ºC, and average minimum into the chromatographic column, in split mode at temperature of 13.26 °C and average relative a ratio of 1:50. The volatile fraction was analyzed humidity of 78.50%. The temperature and relative in an Agilent® 7890A Gas Chromatograph coupled humidity in the greenhouse were recorded with the to an Agilent® 5975C Mass Selective Detector aid of a portable thermo-hygrometer. (MSD) (Agilent Technologies, CA), operated by After 110 days of cultivation (Netto and electron impact ionization using 70 eV, scanning Raffaelli 2004), plants were collected for evaluation mode at a scan rate of 1.0 / s, acquisition mass range of growth. The growth was evaluated through of 40-400 m / z. We used an HP-5MS fused-silica measurement of root length, stem diameter, and capillary column (30 m length x 0.25 mm internal by counting the number of leaves and shoots. The diameter x 0.25 mm film thickness) (California, dry matter production of the aerial part, stem and USA).

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